Liu Yang, Hu Qiang, Wang Xihui
Department of Pharmacy, Linyi Central Hospital, Linyi, Shandong 276400, P.R. China.
Department of Radiology, Linyi Central Hospital, Linyi, Shandong 276400, P.R. China.
Oncol Lett. 2020 Jan;19(1):1024-1030. doi: 10.3892/ol.2019.11175. Epub 2019 Dec 2.
Cisplatin (DDP)-resistance in non-small cell lung carcinoma (NSCLC) severely influences the prognosis of affected patients. This study aims to uncover the potential role of AFAP1-AS1 in DDP-resistant NSCLC and the underlying mechanism. The expression level of AFAP1-AS1 in DDP-resistant NSCLC patients and DDP-resistant A549 cells (A549/DDP) was determined. Proliferative, cell cycle distribution, apoptotic, migratory and invasive changes in A549/DDP cells transfected with si-AFAP1-AS1 were assessed. Western blot analyses were conducted to examine the protein levels of phosphorylated protein kinase B (p-AKT), AKT, E-cadherin, N-cadherin, vimentin and snail in A549/DDP cells. Furthermore, the ubcellular distribution of AFAP1-AS1 was analyzed. Through RNA immunoprecipitation (RIP) assay, the interaction between AFAP1-AS1 and enhancer of zeste homolog 2 (EZH2) was explored. Finally, the regulatory effect of EZH2 on the PI3K/AKT pathway was investigated by western blot analysis. AFAP1-AS1 was upregulated in DDP-resistant NSCLC patients and A549/DDP cells. Transfection with si-AFAP1-AS1 attenuated the proliferative, migratory and invasive abilities, arrested cell cycle in G0/G1 phase, and stimulated apoptosis of A549/DDP cells. Silencing of AFAP1-AS1 upregulated E-cadherin and downregulated N-cadherin, vimentin and snail expression levels. Furthermore, AFAP1-AS1 was verified to interact with EZH2. The relative expression of EZH2 was reduced by transfection of A549/DDP cells with si-AFAP1-AS1. Silencing of EZH2 inhibited the activation of PI3K/AKT pathway. In conclusion, AFAP1-AS1 accelerates the proliferative and metastatic abilities of A549/DDP cells, whereas inhibits the apoptosis of A549/DDP cells, by interacting with EZH2 to activate the PI3K/AKT pathway; thus, inducing DDP resistance in NSCLC.
非小细胞肺癌（NSCLC）中的顺铂（DDP）耐药严重影响患者预后。本研究旨在揭示AFAP1-AS1在DDP耐药NSCLC中的潜在作用及潜在机制。测定了DDP耐药NSCLC患者和DDP耐药A549细胞（A549/DDP）中AFAP1-AS1的表达水平。评估了用si-AFAP1-AS1转染的A549/DDP细胞的增殖、细胞周期分布、凋亡、迁移和侵袭变化。进行蛋白质印迹分析以检测A549/DDP细胞中磷酸化蛋白激酶B（p-AKT）、AKT、E-钙黏蛋白、N-钙黏蛋白、波形蛋白和蜗牛蛋白的水平。此外，分析了AFAP1-AS1的亚细胞分布。通过RNA免疫沉淀（RIP）试验，探索了AFAP1-AS1与zeste同源物2增强子（EZH2）之间的相互作用。最后，通过蛋白质印迹分析研究了EZH2对PI3K/AKT通路的调节作用。AFAP1-AS1在DDP耐药NSCLC患者和A549/DDP细胞中上调。用si-AFAP1-AS1转染可减弱A549/DDP细胞的增殖、迁移和侵袭能力，使细胞周期停滞在G0/G1期，并诱导A549/DDP细胞凋亡。沉默AFAP1-AS1可上调E-钙黏蛋白并下调N-钙黏蛋白、波形蛋白和蜗牛蛋白的表达水平。此外，证实AFAP1-AS1与EZH2相互作用。用si-AFAP1-AS1转染A549/DDP细胞可降低EZH2的相对表达。沉默EZH2可抑制PI3K/AKT通路的激活。总之，AFAP1-AS1通过与EZH2相互作用激活PI3K/AKT通路，加速A549/DDP细胞的增殖和转移能力，同时抑制A549/DDP细胞凋亡；从而诱导NSCLC中的DDP耐药。
